Regeneration of the Lower Urinary Tract in Nonhuman Primates

非人类灵长类动物下尿路的再生

• 批准号: 7641390
• 负责人: James Koudy Williams
• 金额: $ 49.15万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-15 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7641390
• 关键词: Address; Adult; Age; Anatomy; Autologous; Biomedical Engineering; Bladder; Canis familiaris; Cell Survival; Cell Therapy; Cell Transplantation; Cell Volumes; Cells; Characteristics; Clinical; Clinical Trials; Communities; Congenital Abnormality; Data; Deterioration; Electric Stimulation; Engraftment; Excision; Female; Follow-Up Studies; Gene Expression; Genetic; Genitourinary system; Hormones; Human; In Vitro; Injection of therapeutic agent; Injury; Investigation; Ion Channel; Label; LacZ Genes; Location; Lower urinary tract; Macaca fascicularis; Measures; Modeling; Molecular Analysis; Motor Endplate; Muscle; Muscle Cells; Myopathy; Natural regeneration; Organ; Participant; Patients; Pattern; Pelvis; Phenotype; Physiological; Play; Primates; Property; Rattus; Reflex action; Reflex control; Reporter; Research Design; Rest; Role; Skeletal Muscle; Sphincter; Stem cells; Stimulus; Stress Urinary Incontinence; Structure; Subfamily lentivirinae; System; Therapeutic; Time; Transfection; Uncertainty; Urination; Urodynamics; Woman; Work; abstracting; age group; base; electric field; follow-up; functional improvement; in vivo; middle age; muscular structure; nonhuman primate; older women; public health relevance; response; restoration; stressor; success; tissue regeneration; urinary

DESCRIPTION (provided by applicant): The concept of regenerating damaged tissue using selective cell transplantation has been applied clinically for a variety of muscle disorders, including stress urinary incontinence (SUI), which is a manifestation of several kinds of injuries and congenital abnormalities leading to sphincteric muscle deficiency (SMD). Results of recent clinical trials using autologous muscle-derived progenitor cells (MPCs) to treat SMD in women are promising. However, it remains unclear whether injected MPCs play a passive (bulking), or active (cell integration) role in sphincter regeneration and recent concern has been raised about the short- and long-term success rates of these clinical trials. Repeated clinical trials will take time and still lack a mechanistic explanation for their results. We propose to address these two issues in a study designed to evaluate the role of MPCs as active participants in restoration of reflex control over micturition using a cynomolgus monkey (Cyno) model of injury- induced SMD, where measures of success can be achieved in relatively few years. Two findings from our lab (one in rats and one in dogs) support the working hypothesis that injected MPCs integrate and play an active role in sphincter regeneration of females. Based on the current uncertainties associated with this therapeutic approach and the need for results that are more directly translatable to human beings, we propose to extend our studies to adult female Cyno's - at the age equivalent of 45-50 year-old women - whose genetic, anatomic, physiologic, postural, intra-pelvic bladder location, age and hormone-related changes in the genitourinary system, expose the urinary sphincter to structural and pharmacologic stressors common to middle-aged women. The specific aims are: 1. To establish a Cyno model of injury-induced SMD. This will be achieved by applying our canine model of sphincter muscle removal to adult female Cyno's and then measuring longitudinal changes in urinary sphincter structure (histological and molecular analysis of engraftment characteristics such as: sphincter content of labeled cells, injected cell viability, formation of functional motor endplates and gene expression data) and function (resting and reflex urodynamics, responses to pharmacologic and electric field stimulation) at 3mo., 6mo., 12mo., and 24 mo. post injury ( this will be equivalent to a 6 year follow-up study in human beings); and 2. To measure the effects of MPC cell therapy in the Cyno model of injury-induced SMD. First, we will optimize the culture system to produce sufficient numbers of functional lenti-LacZ transfected Cyno-derived MPCs for use in the treatment of SMD and then inject the labeled MPCs into the damaged sphincter muscle and then measure the longitudinal (same time- points as SA1) effects of these cells on sphincter function and structure. It is anticipated that the results obtained from this primate model will be directly translatable to women of this age-group, to other patients with SMD and to regeneration of other muscle tissues and muscle cell-based bioengineered organs. PUBLIC HEALTH RELEVANCE: An unanswered question about cell therapy for sphincter muscle deficiency (SMD) -associated stress urinary incontinence is whether the injected cells play a passive, or an active role in clinical improvement. The proposed studies will utilize adult female nonhuman primates to measure the longitudinal effects of muscle progenitor cells on both structural and functional changes in an injury model of SMD. We anticipate that longitudinal and parallel assessments of structure (MPC viability, functionality, and contribution to motor endplate formation) and function (resting and reflex urodynamic measures) will help determine the role (active or passive) of MPC cell therapy in restoration of a functional sphincter.

描述（由申请人提供）：使用选择性细胞移植再生受损组织的概念已在临床上应用于多种肌肉疾病，包括应激性尿失禁（SUI），这是几种损伤和先天性异常导致括约肌缺陷（SMD）的表现。最近使用自体肌源性祖细胞（MPCs）治疗女性SMD的临床试验结果很有希望。然而，目前尚不清楚注射的MPCs在括约肌再生中是否起被动（增大）或主动（细胞整合）作用，最近人们对这些临床试验的短期和长期成功率提出了关注。重复的临床试验需要时间，而且仍然缺乏对其结果的机制解释。我们建议在一项研究中解决这两个问题，该研究旨在利用食蟹猴（Cyno）损伤性SMD模型来评估MPCs作为积极参与者在恢复对排尿的反射控制中的作用，在相对较短的时间内可以取得成功。我们实验室的两个发现（一个在大鼠和一个在狗身上）支持了注射MPCs整合并在雌性括约肌再生中发挥积极作用的工作假设。基于目前与这种治疗方法相关的不确定性以及对更直接适用于人类的结果的需求，我们建议将我们的研究扩展到成年女性Cyno -年龄相当于45-50岁的女性-其遗传，解剖，生理，体位，盆腔内膀胱位置，年龄和泌尿生殖系统激素相关的变化。将尿括约肌暴露在中年妇女常见的结构和药理学压力下。具体目标是：1。建立损伤性SMD的Cyno模型。这将通过将我们的犬括约肌切除模型应用于成年雌性Cyno，然后在3个月时测量尿括约肌结构的纵向变化（对植入特征的组织学和分子分析，如：标记细胞的括约肌含量、注射细胞活力、功能性运动终板的形成和基因表达数据）和功能（静息和反射尿动力学、对药物和电场刺激的反应）。6 mo。12个月,。，伤后24小时（相当于对人类进行6年的随访研究）；和2。观察MPC细胞治疗对损伤性SMD Cyno模型的影响。首先，我们将优化培养系统，以产生足够数量的功能性lentil - lacz转染cyno衍生的MPCs用于SMD的治疗，然后将标记的MPCs注射到受损的括约肌中，然后测量这些细胞对括约肌功能和结构的纵向（与SA1相同的时间点）影响。预计从该灵长类动物模型获得的结果将直接转化为该年龄组的女性，其他SMD患者以及其他肌肉组织和基于肌肉细胞的生物工程器官的再生。公共卫生相关性：关于细胞治疗括约肌缺陷（SMD）相关性应激性尿失禁的一个悬而未决的问题是，注射细胞在临床改善中是被动的还是主动的作用。该研究将利用成年雌性非人灵长类动物来测量肌肉祖细胞对SMD损伤模型中结构和功能变化的纵向影响。我们预计，纵向和平行评估结构（MPC活力、功能和对运动终板形成的贡献）和功能（静息和反射尿动力学测量）将有助于确定MPC细胞治疗在恢复功能性括约肌中的作用（主动或被动）。